JP2007037422A - Culture treatment apparatus and culture treatment method - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To prevent the occurrence of troubles such as the contamination of dusts or the like in a culture container, or the adhesion or contamination of the droplet or aerosol of liquids in various containers including the culture container to various other containers. <P>SOLUTION: This culture treatment apparatus 1 for applying prescribed treatments to cells received in the culture container 9 is characterized by comprising a small space 4 divided from the outside, air lock type pass boxes 14, 15 capable of putting in and out various containers such as the culture container 9, a chemical liquid container 7 and a serum container 8 capable of being closed or opened with lids 7a to 10a, 12a, in the small space 4, a conveying mechanism 21 capable of conveying the various containers on the inside and outside of the small space 4 through the pass boxes 14, 15, and a controller 31 for inhibiting the introduction of various containers into the small space 4, when a cleanliness factor measured with cleanliness factor sensors 18 does not satisfy a prescribed cleanliness. Closing or opening mechanisms 19 for closing or opening the lids of the various containers, a dispense device 20 capable of sucking/dispensing chemical liquids or the like from the introduced various containers, and the cleanliness factor sensors 18 for measuring cleanliness factors in the small space 4 are disposed in the small space 4. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a culture treatment apparatus and a culture treatment method used in an automatic culture apparatus.

As a conventional automatic culture apparatus, an apparatus including a fixed storage shelf that can store a plurality of culture vessels and a conveying means that can move horizontally, vertically, and rotationally is known (see, for example, Patent Document 1). .)
In this automatic culture apparatus, after the culture container is taken out of the incubator by the transfer robot, the lid of the culture container is removed by the lid open robot, and the medium in the culture container is discharged and injected by the medium exchange robot. ing. After completion of the medium exchange, the lid of the culture container is again covered by the lid open robot, and the culture container is accommodated in the stacker in the incubator by the transport robot.

In the automatic culture apparatus described in Patent Document 1, the inside of the stacker that accommodates the culture vessel is managed so that culture conditions such as temperature, humidity, and carbon dioxide concentration are kept constant. On the other hand, a space for removing the lid of the culture container or performing various treatments such as medium exchange on the opened culture container is arranged outside the stacker.
JP 2002-262856 A

  However, since various mechanical devices such as robots and transfer conveyors are arranged in the space outside the stacker, it is an environment in which dust tends to float, and it is necessary to reduce the floating dust by some means. If not dealt with, it is conceivable that when the lid of the culture vessel is removed, dust or the like is mixed into the culture vessel.

  In addition, when a liquid such as a medium is supplied into the culture container, the supplied medium and the cells in the culture container may be splashed and scattered. Furthermore, when the lids of various containers such as a culture container, a drug solution container, or a serum container are opened, the liquid in these various containers may be aerosolized and float in the space.

  In such a case, in the automatic culture apparatus of Patent Document 1, it is conceivable that the scattered droplets and the floating aerosol adhere to other various containers or are mixed into the various containers. In particular, the cells in the culture container and the serum in the serum container are unique to each specimen, and when handling multiple specimens with the same automatic culture equipment, take care not to cause cross-contamination problems. Must.

  The present invention has been made in view of the above-mentioned circumstances, and it is possible to mix dust and the like in the culture container, and liquid droplets and aerosols in various containers including the culture container may adhere to other various containers. An object of the present invention is to provide a culture treatment apparatus and a culture treatment method capable of preventing the occurrence of inconveniences such as mixing.

In order to achieve the above object, the present invention provides the following means.
In order to achieve the above object, a culture treatment apparatus for performing predetermined treatment on cells contained in a culture vessel, which can be opened and closed by a lid in a small space partitioned from the outside and the small space An air-lock type pass box capable of taking in and out various containers such as a culture container, a drug solution container and a serum container, and a transport mechanism capable of transporting various containers into and out of the small space via the pass box, An open / close mechanism that opens and closes the lids of the various containers, a dispensing device that can suck and dispense chemicals and the like from the various containers introduced, and a cleanliness sensor that measures the cleanliness of the small space Provided is a culture treatment apparatus provided with a control unit that is arranged and prohibits introduction of various containers into a small space when the cleanliness measured by the cleanliness sensor does not satisfy a predetermined cleanliness.

  According to the present invention, when a treatment for supplying a liquid such as a chemical solution, serum, or culture medium is performed in a culture container, first, various containers such as a chemical solution container and a serum container containing the liquid to be supplied are provided with a pass box. Through a small space. Various containers are transported into the small space by the operation of the transport mechanism. Since the pass box is configured as an air lock type, the atmosphere outside the small space can be brought into the small space, or the atmosphere inside the small space can be kept from being taken out outside the small space.

  When various containers are introduced into the small space, the lid of the introduced various containers is opened by the operation of the opening / closing mechanism arranged in the small space, and the dispensing device removes the necessary amount of liquid in the various containers. Suction. In this conception, the lids of the various containers are closed again by the operation of the opening / closing mechanism, and the various containers are taken out of the small space through the pass box by the operation of the transport container. Thereafter, the culture container is introduced into the small space through the pass box, the lid is opened by the operation of the opening / closing mechanism, and the retained liquid is supplied into the culture container by the operation of the dispensing device. The

  In this case, according to the present invention, even if splashes occur due to the supply of liquid into the culture container, other containers are not disposed in the small space, so the splashes adhere to other containers, Generation | occurrence | production of the problem which mixes in these containers can be prevented beforehand. As a result, occurrence of cross contamination can be prevented.

  Further, according to the present invention, the cleanliness sensor monitors the cleanliness in the small space, and when the predetermined cleanliness is not satisfied, the operation of the control unit prohibits the introduction of various containers into the small space. Therefore, when the lids of various containers are opened, it is possible to prevent other containers from being introduced into the small space in a state where the liquid inside becomes an aerosol and is suspended in a large amount in the small space. As a result, it is possible to prevent the occurrence of cross-contamination due to aerosol through other containers.

In the said invention, the said conveyance mechanism is each arrange | positioned in the said pass box and the said small space, It is provided with the some belt conveyor which delivers the various containers to convey, The said belt is provided in the bottom part of the said pass box and the said small space. It is preferable that a disinfectant bath for immersing a part of the conveyor is provided.
By configuring in this way, even if splashes generated by supplying the liquid into the culture vessel adhere to the belt conveyor, the entire belt conveyor can be passed through the disinfecting liquid tank by operating the belt conveyor. Further, it is possible to prevent the droplets adhering to the belt conveyor from adhering to various containers and being taken out of the small space.

  The present invention also relates to a culture treatment method for supplying a liquid contained in various containers such as a drug solution container and a serum container to cells contained in the culture container, wherein the small space has a controlled cleanliness. After introducing various containers, opening the lids of the various containers introduced, sucking and holding the liquid inside the various containers, closing the lids of the various containers and taking them out of the small space, A culture treatment method is provided in which a culture vessel is introduced, a lid of the introduced culture vessel is opened, and liquids in various held containers are injected into the culture vessel.

  According to the present invention, when the liquid is injected into the culture container, since the various containers that contain the liquid are not arranged in the same small space, the liquid droplets from the culture container adhere to the various containers. It is possible to prevent inconveniences such as mixing into various containers.

In the said invention, after confirming that the cleanliness in a small space is more than predetermined cleanliness, it is preferable to introduce a culture container and various containers into the small space.
In this way, when the culture vessel or various containers are opened, it is possible to introduce other culture vessels or various containers into the small space in a state where a liquid is generated as an aerosol and a large amount is suspended in the small space. Is prevented. As a result, it is possible to prevent the occurrence of cross-contamination due to aerosol through other containers.

  According to the present invention, it is possible to prevent the occurrence of inconveniences such as contamination of dust or the like in the culture container, liquid droplets in various containers including the culture container, and aerosol adhering to or mixing in other various containers. There is an effect that can be.

Hereinafter, the culture treatment apparatus 1 which concerns on one Embodiment of this invention is demonstrated with reference to FIGS.
As shown in FIG. 1, the culture treatment apparatus 1 according to the present embodiment has a second space (small space) 4 further partitioned by a partition wall 3 in a first space 2 partitioned from the outside. Is provided. Outside the second space 4 in the first space 2, a chip box 6 containing a plurality of disposable chips 5, a chemical solution container 7 containing various chemical solutions, a serum container 8 containing serum, A culture container 9 containing cells, a waste container 10 for discarding a used medium aspirated from the culture container 9, a centrifuge 11, a centrifuge container 12 used at the time of centrifugation by the centrifuge 11, and the like. It is arranged. The first space 2 is provided with a handling robot 13 for handling these various containers 7, 8, 10, 12 and the chip box 6.

Various containers such as the culture container 9, the drug solution container 7, the serum container 8, the disposal container 10, and the centrifuge container 12 are provided to be opened and closed by lid bodies 7 a to 10 a and 12 a, respectively. Examples of the chemical solution stored in the chemical solution container 7 include a proteolytic enzyme such as trypsin that peels off the cells in the culture container 9, a growth factor such as a cytokine that grows the cells during the culture, and dexamethasone that differentiates the cells. Examples include differentiation inducers, antibiotics such as penicillin antibiotics, hormones such as estrogen, and nutrients such as vitamins. The serum container 8 also has a MEM (Minimal Essential) that constitutes a medium necessary for culturing cells.
Medium: Minimum essential medium), serum such as DMEM (Dulbecco's Modified Eagle Medium), FBS (Fetal Bovine Serum) and human serum are contained.

The drug solution container 7 and the serum container 8 are accommodated in a cool box (not shown), and are taken out into the first space 2 as necessary.
The culture vessel 9 is cultivated in an incubator (not shown), taken out from the incubator as needed, for example, at the time of medium exchange or passage, and is transported into the first space 2.

  The second space 4 is provided with pass boxes 14 and 15 that allow various containers 7 to 10 and 12 and a chip box 6 to be taken in and out of the second space 4. In the example shown in FIG. 1, the pass boxes 14 and 15 are provided at two locations on the partition wall 3 facing the second space 4. Each of the pass boxes 14 and 15 includes shutters 14a, 14b, 15a, and 15b that can be opened and closed on both the entrance side and the exit side to the second space 4. The second space 4 and each of the pass boxes 14 and 15 are provided with an air cleaner 16 for purifying internal air. Further, a sterilization lamp 17 is attached in the second space 4. A particle counter 18 that detects the number of dust in the second space 4 is provided in the second space 4 and the pass boxes 14 and 15.

In addition, the second space 4 temporarily sucks the liquid in the containers 7 to 10 and 12 by sucking the liquid in the containers 7 to 10 and 12 and the opening / closing mechanism 19 that can open and close the lids 7a to 10a and 12a of the containers 7 to 10 and 12. And a dispensing device 20 that can be held and discharged if necessary.
In the pass boxes 14 and 15 and the second space 4, a transport mechanism 21 that receives and transports the various containers 7 to 10 and 12 and the chip box 6 transported from the first space 2 is provided. Is provided. In the example illustrated in FIG. 2, the transport mechanism 21 includes a plurality of belt conveyors 22 arranged in series. Each belt conveyor 22 is driven synchronously, and the various containers 7 to 10, 12 and the like to be conveyed are smoothly delivered from one belt conveyor 22 to another belt conveyor 22.

  In the pass boxes 14 and 15 and the second space 4, a disinfecting liquid tank 24 capable of storing a disinfecting liquid such as ethanol 23 is provided at the bottom. Each disinfecting liquid tank 24 stores ethanol 23 to such an extent that a part of the belt constituting the belt conveyor 22 is immersed. Thereby, when the belt conveyor 22 is driven and the belt is moved, the belt is sterilized by being sequentially passed through the disinfecting liquid tank 24. Further, in each of the pass boxes 14 and 15, an ethanol shower 25 for injecting the mist-like ethanol 23 into the various containers 7 to 10 and 12 passing through the pass boxes 14 and 15 is attached.

  Further, a part of the belt conveyor 22 'in FIG. 2 is equipped with a swing mechanism (not shown). As a result, the belt conveyor 22 ′ is swung around the horizontal axis while the culture vessel 9 is mounted, so that the mounted culture vessel 9 can be swung to stir the liquid inside. It has become.

  The opening / closing mechanism 19 includes a chuck 26 that holds the lids 7a to 10a and 12a of the various containers 7 to 10 and 12 conveyed by the conveyance mechanism 21, and a first moving mechanism that moves the chuck 26 in the vertical direction. 27 and a rotation mechanism 28 such as a motor for rotating the chuck 26 holding the lids 7a to 10a and 12a around the vertical axis. In the example shown in FIG. 1, an opening / closing mechanism 19 that opens and closes the lid 9 a of the culture vessel 9 that has been transported into the second space 4 via one pass box 14 and another pass box 15. Two open / close mechanisms 19 for opening and closing the lids 7a, 8a, 10a, 12a of the other various containers 7, 8, 10, 12 that have been transported into the second space 4 are provided.

  The dispensing device 20 includes a nozzle 29 that detachably attaches the chip 5 accommodated in the chip box 6 and a second moving mechanism 30 that moves the nozzle 29 up and down and horizontally. The nozzle 29 is connected to a dispensing control device (not shown), and sucks and holds the liquid in the tip 5 attached to the tip by suction, and can discharge the liquid in the tip 5 by discharging. It has become.

  Further, each of the pass boxes 14 and 15, the belt conveyor 22 constituting the transport mechanism 21, the opening / closing mechanism 19, the dispensing device 20, and the particle counter 18 are all connected to the control unit 31. The control unit 31 receives the detection result of the particle counter 18 and controls the shutters 14a, 14b, 15a, 15b, the ethanol shower 25, the transport mechanism 21, the opening / closing mechanism 19, and the dispensing device 20 of the pass boxes 14, 15. It has become.

  Specifically, as a result of detection by the particle counter 18, when it is determined that the cleanliness in the second space 4 is lower than the class 100 (that is, the cleanliness is low), the passbox 14 from the second space 4 is used. , 15 is allowed to be discharged into the second space 4 from the inside of the pass box 14, 15, while the culture container 9 and the various containers 7, 8, 10, 12 are allowed to be discharged. The introduction of 10, 12 and the chip box 6 is prohibited.

  Further, when any of the containers 7 to 10 and 12 is accommodated in the second space 4 in a state of being closed by the lids 7a to 10a and 12a, the second counter 4 detects the second result. When it is determined that the cleanliness in the space 4 is lower than the class 100 (that is, the cleanliness is low), the control unit 31 prohibits the opening / closing mechanism 19 from opening the lids 7a to 10a and 12a.

The operation of the culture treatment apparatus 1 according to the present embodiment configured as described above will be described below.
Here, the case where the subculture process which replaces | exchanges the culture container 9 is performed with respect to the cell cultured in the culture container 9 using the culture processing apparatus 1 which concerns on this embodiment is demonstrated.

  First, the shutters 14a, 14b, 15a, 15b of the two pass boxes 14, 15 provided in the second space 4 are closed, and the inside of the second space 4 and the inside of the pass boxes 14, 15 are sealed. Each air cleaner 16 is operating. The control unit 31 monitors the output of the particle counter 18 in the pass boxes 14 and 15 to determine whether the cleanliness inside the pass boxes 14 and 15 is class 100 or higher.

When the cleanliness in the pass boxes 14 and 15 is less than class 100, the control unit 31 opens the shutters (exit side shutters) 14b and 15b between the pass boxes 14 and 15 and the second space 4. It is prohibited and waits until the cleanliness in the pass boxes 14 and 15 becomes class 100 or higher by the operation of the air cleaner 16 or the operation of the ethanol shower 25.
When the cleanliness in the pass boxes 14 and 15 becomes class 100 or higher, as shown in FIG. 2, the control unit 31 opens the entrance-side shutters 14a and 15a of the pass boxes 14 and 15 for handling. The chip box 6 is mounted on the belt conveyor 22 following the one pass box 15 by the operation of the robot 13. A culture vessel 9 containing cells is introduced into the other pass box 14.

  When the chip box 6 and the culture vessel 9 are introduced into the pass boxes 14 and 15 by the operation of the belt conveyor 22, the entrance side shutters 14a and 15a are closed, and the pass boxes 14 and 15 are sealed. In this state, the air purifier 16 of the pass boxes 14 and 15 is activated again, and the control unit 31 causes the output side of the particle counter 18 inside the pass boxes 14 and 15 to exit until the cleanliness reaches a class 100 or higher. Opening of the shutters 14b and 15b is prohibited.

At the same time, the control unit 31 monitors the cleanliness in the second space 4 based on the output of the particle counter 18 in the second space 4. If the cleanliness is less than class 100, the control unit 31 is monitored. Prohibits the opening of the exit side shutters 14b, 15b of the pass boxes 14, 15.
When the cleanliness in the second space 4 becomes class 100 or higher, the control unit 31 first opens the exit-side shutter 15b of the pass box 15 in which the chip box 6 is accommodated, and the belt conveyor 22 By the operation of, the chip box 6 in the pass box 15 is introduced into the second space 4. When the chip box 6 is introduced into the second space 4, the control unit 31 closes the outlet side shutter 15b. Further, the control unit 31 further drives the belt conveyor 22 to move the chip box 6 to a position where a new chip 5 is disposed below the nozzle 29 of the dispensing device 20 and stops the chip box 6 on the spot.

In this state, the control unit 31 operates the second moving mechanism 30 of the dispensing device 20 to lower the nozzle 29, so that the new chip 5 in the chip box 6 is moved to the nozzle as shown in FIG. After mounting, the nozzle 29 with the tip 5 is raised to a predetermined position.
Thereafter, the exit-side shutter 15b of the pass box 15 is opened, and the operation of the belt conveyor 22 causes the chip box 6 in the second space 4 to be retracted into the pass box 15, and the exit-side shutter 15b is closed. . Even in this case, the opening of the entrance-side shutter 15a is prohibited until the cleanliness in the pass box 15 becomes more than class 100 by the operation of the air cleaner 16, and the entrance-side shutter 15a is opened after reaching the class 100 or more. Then, the chip box 6 is taken out into the first space 2.
The waste container 10 to be introduced into the second space 4 is introduced into the pass box 15 that has been emptied after the chip box 6 is taken out, and is placed in a standby state in the pass box 15.

Next, the control unit 31 monitors the cleanliness in the second space 4 based on the output of the particle counter 18 in the second space 4, and when the cleanness is less than class 100, the pass boxes 14 and 15 The opening of the exit side shutters 14a and 15a is prohibited.
When the cleanliness in the second space 4 is equal to or higher than class 100, the control unit 31 opens the outlet side shutter 14b of the pass box 14 in which the culture vessel 9 is accommodated, and operates the belt conveyor 22. Thus, the culture container 9 in the pass box 14 is introduced into the second space 4. When the culture container 9 is introduced into the second space 4, the control unit 31 closes the outlet side shutter 14b. Further, the control unit 31 further drives the belt conveyor 22 to move the culture vessel 9 below the opening / closing mechanism 19 and stop it on the spot.

Even in this state, the control unit 31 monitors the cleanliness in the second space 4, and prohibits the opening / closing operation of the lid 9a by the opening / closing mechanism 19 when the cleanness is less than class 100.
When the cleanliness in the second space 4 becomes class 100 or higher, the control unit 31 operates the first moving mechanism 27 of the opening / closing mechanism 19 so that the chuck 26 is attached to the lid body 9a of the culture vessel 9. The lid body 9a is gripped by operating the chuck 26, and the lid body 9a is rotated and opened by the operation of the rotation mechanism 28. Then, as shown in FIG. 4, the second moving mechanism 30 of the dispensing device 20 is operated to insert the nozzle 29 with the tip 5 into the culture vessel 9, and the medium in the culture vessel 9 is put into the tip 5. To suck.

  After the nozzle 29 with the tip 5 that has sucked the culture medium is retracted from the culture container 9, the culture container 9 is closed by the lid 9a by the operation of the opening / closing mechanism 19 again. Then, the exit side shutter 14b of the pass box 14 is opened, the belt conveyor 22 is operated, the culture vessel 9 is retracted into the pass box 14, and the exit side shutter 14b is closed.

Even in this state, the control unit 31 monitors the cleanliness in the second space 4 and prohibits the opening of the exit side shutters 14b and 15b of the pass boxes 14 and 15 when the cleanness is less than class 100. To do.
When the cleanliness in the second space 4 is equal to or higher than class 100, the control unit 31 opens the outlet side shutter 15b of the pass box 15 in which the waste container 10 is accommodated, and operates the belt conveyor 22. Thus, the waste container 10 in the pass box 15 is introduced into the second space 4.

  When the waste container 10 is introduced into the second space 4, the control unit 31 closes the outlet side shutter 15b. Further, the control unit 31 further drives the belt conveyor 22 to move the waste container 10 below the opening / closing mechanism 19 and stop it on the spot.

  The control unit 31 operates the first moving mechanism 27 of the opening / closing mechanism 19 to place the chuck 26 at the position of the lid 10a of the waste container 10, operates the chuck 26 to grip the lid 10a, and rotates the rotation mechanism. By the operation of 28, the lid 10a is rotated and opened. Then, the second moving mechanism 30 of the dispensing device 20 is operated to insert the nozzle 29 with the tip 5 containing the culture medium into the waste container 10 and discharge the culture medium into the waste container 10.

  When the culture medium is discharged into the waste container 10, the opening / closing mechanism 19 is actuated again, and the lid 10a of the waste container 10 is closed. Then, the exit-side shutter 15b of the pass box 15 is opened, and the waste container 10 containing the waste medium in the second space 4 is retracted into the pass box 15 by the operation of the belt conveyor 22, and the exit-side shutter 15b. Is closed. Even in this case, the opening of the entrance-side shutter 15a is prohibited until the cleanliness in the pass box 15 becomes more than class 100 by the operation of the air cleaner 16, and the entrance-side shutter 15a is opened after reaching the class 100 or more. Then, the waste container 10 is taken out into the first space 2.

Next, the tip 5 of the dispensing device 20 is replaced. The used tip 5 attached to the nozzle 29 of the dispensing device 20 is removed from the nozzle 29 by operating the second moving mechanism 30 and hooking it on a hook (not shown). The removed chip 5 is collected in a chip collection container (not shown).
Thereafter, a new chip 5 is supplied into the second space 4. The supply of the chip 5 is as described above.

  After the new chip 5 is attached to the nozzle 29, the drug solution container 7 containing a proteolytic enzyme such as trypsin is introduced into the pass box 15. In this case as well, when the cleanliness in the pass box 15 is equal to or higher than class 100, the control unit 31 opens the entrance-side shutter 15a of the pass box 15 and operates the handling robot 13. The chemical container 7 is mounted on the belt conveyor 22 following the one pass box 15.

  When the chemical container 7 is introduced into the pass box 15 by the operation of the belt conveyor 22, the entrance side shutter 15a is closed and the pass box 15 is sealed. In this state, the air purifier 16 of the pass box 15 is activated again, and the control unit 31 opens the outlet side shutter 15b until the cleanliness becomes class 100 or higher by the output of the particle counter 18 inside the pass box 15. Is prohibited.

At the same time, the control unit 31 monitors the cleanliness in the second space 4 based on the output of the particle counter 18 in the second space 4, and if the cleanness is less than class 100, the pass box 14 , 15 are prohibited from opening the outlet side shutters 14b, 15b.
When the cleanliness in the second space 4 is equal to or higher than class 100, the control unit 31 opens the exit-side shutter 15b of the pass box 15 in which the chemical solution container 7 is accommodated, and the belt conveyor 22 operates. Thus, the chemical solution container 7 in the pass box 15 is introduced into the second space 4. When the chemical liquid container 7 is introduced into the second space 4, the control unit 31 closes the outlet shutter 15b. Further, the control unit 31 further drives the belt conveyor 22 to move the chemical solution container 7 below the opening / closing mechanism 19 and stop it on the spot.

  The control unit 31 operates the first moving mechanism 27 of the opening / closing mechanism 19 to place the chuck 26 at the position of the lid body 7a of the chemical liquid container 7, operates the chuck 26 to grip the lid body 7a, and rotates the rotation mechanism. By the operation of 28, the lid 7a is rotated and opened. Then, as shown in FIG. 6, the second moving mechanism 27 of the dispensing device 20 is operated to insert the nozzle 29 with the new tip 5 attached into the chemical solution container 7, and trypsin in the chemical solution container 7. Is sucked into the tip 5.

After the nozzle 29 with the tip 5 that has sucked in trypsin is withdrawn from the chemical container 7, the lid 7a closes the chemical container 7 again by the operation of the opening and closing mechanism 19, and the exit side shutter 15b of the pass box 15 is opened. The belt conveyor 22 is operated, the chemical container 7 is retracted into the pass box 15, and the outlet shutter 15b is closed.
Even in this state, the control unit 31 monitors the cleanliness in the second space 4 and, when the cleanness is less than class 100, prohibits the opening of the exit side shutters 14a and 15a of the pass boxes 14 and 15. To do.

  When the cleanliness in the second space 4 is equal to or higher than class 100, the control unit 31 opens the outlet side shutter 14b of the pass box 14 in which the culture vessel 9 is accommodated, and operates the belt conveyor 22. Thus, the culture container 9 in the pass box 14 is introduced into the second space 4. When the culture container 9 is introduced into the second space 4, the control unit 31 closes the outlet side shutter 14b. Further, the control unit 31 further drives the belt conveyor 22 to move the culture vessel 9 below the opening / closing mechanism 19 and stop it on the spot.

Even in this state, the control unit 31 monitors the cleanliness in the second space 4, and when the cleanness is less than class 100, the control unit 31 prohibits the opening / closing operation of the lid 9 a by the opening / closing mechanism 19. To do.
When the cleanliness in the second space 4 is equal to or higher than class 100, the control unit 31 operates the first moving mechanism 27 of the opening / closing mechanism 19 to open the lid body 9a of the culture vessel 9. . Then, the second moving mechanism 30 of the dispensing apparatus 20 is operated to insert the nozzle 29 with the tip 5 into the culture vessel 9, and the trypsin sucked into the tip 5 is released into the culture vessel 9.

  The lid 9a of the culture container 9 is attached to the culture container 9 into which trypsin has been injected in the same manner as described above. Then, as shown in FIG. 7, the culture vessel 9 is moved onto a belt conveyor 22 ′ having a rocking mechanism, and the culture vessel 9 is rocked to stir trypsin to promote cell detachment. After the predetermined time has elapsed, when the cells are detached from the bottom surface of the culture container 9, the lid body 9 a of the culture container 9 is removed again by the opening / closing mechanism 19, and the cell suspension formed in the culture container 9 is put into the chip 5. Suction.

  The culture vessel 9 evacuated by sucking the internal cell suspension is attached to the lid 9a by the opening / closing mechanism 19 and then discharged to the outside of the second space 4 via the pass box 14, Discarded. On the other hand, in the same pass box 14, a new culture vessel 9 is prepared and waits until the next processing is performed.

  Next, in the second space 4, as shown in FIG. 8, the centrifuge container 12 is introduced in the same procedure as in the case of the waste container 10, and the cell suspension in the chip 5 is placed in the centrifuge container 12. The liquid is poured and moved from the second space 4 to the first space 2 in a closed state. Then, the cells are separated by being put into a centrifuge disposed in the first space 2 and then introduced into the second space 4 again and separated by the nozzle 29 with the chip 5. Cells are aspirated into the chip 5. Since the separated cells sink below the centrifuge container 12, only the cells can be sucked into the chip 5 simply by inserting the tip of the chip 5 to the bottom surface of the centrifuge container 12.

Thereafter, the centrifuge container 12 is closed by the lid 12 a and then discharged out of the second space 4 in the same manner as the waste container 10.
After the centrifuge container 12 is discharged out of the second space 4, the outlet side shutter 14 b of the pass box 14 is opened, and a new culture container 9 is introduced into the second space 4. When the lid 9 a of the culture container 9 is opened by the opening / closing mechanism 19, the cells in the chip 5 are seeded in the new culture container 9 by the dispensing device 20.

  The culture container 9 seeded with the cells is once closed and retracted into the pass box 14. Then, after the tip 5 at the tip of the nozzle 29 is replaced in the same manner as described above, the serum container 8 is introduced into the second space 4 via the pass box 15. Even in this case, when the cleanliness in the second space 4 is lower than the class 100, the introduction of the serum container 8 is prohibited, and when the cleanliness is higher than the class 100, the outlet side shutter 14b of the pass box 14 is opened and the serum is released. A container 8 is introduced into the second space 4.

Then, the cleanliness in the second space 4 is checked again, and the lid 8a is opened when the cleanliness is class 100 or higher.
A new tip 5 is inserted into the serum container 8 with the lid 8 a opened, and a predetermined amount of serum is sucked into the tip 5 by the nozzle 29 of the dispensing device 20.

  After the tip 5 of the dispensing device 20 is retracted from the serum container 8, the opening and closing mechanism 19 is operated to close the serum container 8 by the lid 8 a, open the exit-side shutter 15 b of the pass box 15, and the belt conveyor 22. Is operated to retract the serum container 8 into the pass box 15 and close the outlet shutter 15b. The serum container 8 is discharged from the pass box 15 to the first space 2 after the cleanliness in the pass box 15 reaches class 100 or higher.

  After the serum container 8 is retracted in the pass box 15, the cleanliness in the second space 4 is checked. If the cleanliness is class 100 or higher, the culture container 9 is transferred from the other pass box 14 to the second. Is introduced into the space 4. When the cleanliness is less than or equal to class 100, the outlet shutter 14b is kept closed.

  Thereafter, in the same manner as described above, the lid body 9a of the culture container 9 is opened, the serum held in the chip 5 is injected into the culture container 9, and the lid body 9a is closed again. The culture vessel 9 with the lid 9a closed is retracted to the pass box 14, and after the chip 5 is removed from the nozzle 29 in the same manner as described above, the cleanliness in the second space 4 is class 100 or higher. On the condition that the chip box 6 holding the new chip 5 is introduced into the second space 4, the new chip 5 is mounted on the nozzle 29. The chip box 6 that has finished supplying new chips 5 is discharged to the first space 2 via the pass box 14.

  Next, a chemical solution container 7 containing a necessary chemical solution such as a growth factor is introduced into the second space 4 through the pass box 15. Even in this case, the introduction of the chemical container 7 is permitted on the condition that the cleanliness in the second space 4 is class 100 or higher. In the same manner as described above, the lid 7a is opened with respect to the drug solution container 7, the drug solution is sucked and held by the dispensing device 20, and is retracted into the pass box 15 after the lid 7a is closed. Thereafter, the chemical container 7 is discharged into the first space 2 on the condition that the cleanliness in the pass box 15 is class 100 or higher.

After the chemical container 7 is retracted in the pass box 15, the introduction of the culture container 9 into the second space 4 is permitted on condition that the cleanliness in the second space 4 is class 100 or higher. Then, the lid body 9a is opened, and the chemical solution is introduced into the culture container 9 by the dispensing device 20.
In this way, the culture vessel 9 into which the necessary chemical solution has been injected is discharged into the first space 2 via the pass box 14 and returned to the incubator (not shown) disposed outside the first space 2. Then, the culture process is continued.

  As described above, according to the culture treatment apparatus 1 according to the present embodiment, the cell suspension, serum, drug solution, and the like are supplied from the chip 5 attached to the nozzle 29 to the culture container 9 in the second space 4. In this case, various liquid droplets are generated. At this time, since the other containers 7, 8, 10, 12 and the like are not arranged in the second space 4, the droplets are not transferred to the other containers 7, 8. , 10, 12 and the like, and the inconvenience of being mixed into these containers 7, 8, 10, 12 can be prevented.

Moreover, according to the culture treatment apparatus 1 according to the present embodiment, when the various containers 7 to 10 and 12 are opened in the second space 4, the liquid inside becomes an aerosol in the second space 4. Although a large amount of floating is considered, the cleanness in the second space 4 is monitored by the particle counter 18 disposed in the second space 4, and when the predetermined cleanness is not satisfied, Since the introduction of the various containers 7 to 10 and 12 into the second space 4 is prohibited, it is possible to prevent the liquid in the form of aerosol from adhering to the other containers 7 to 10 and 12.
As a result, it is possible to prevent the occurrence of cross contamination via droplets or aerosols.

  Moreover, according to this embodiment, the pass boxes 14 and 15 provided with the two shutters 14a, 14b, 15a, and 16b in an air-locked manner are provided at the connecting portion between the first space 2 and the second space 4. Since the various containers 7 to 10 and 12 are put into and out of the second space 4 through the pass boxes 14 and 15, dust or the like in the first space 2 is brought into the second space 4. In addition, it is possible to prevent liquid splashes and aerosols generated in the second space 4 from being released into the first space 2.

  Furthermore, since a part of the belt conveyor 22 that conveys the various containers 7 to 10, 12, etc. between the first space 2 and the second space 4 is immersed in the disinfecting liquid 23, the driving of the belt conveyor 22 is performed. Thus, the belt conveyor 22 is driven while being passed through the disinfecting liquid 23 each time. Therefore, even if liquid droplets or aerosols in the various containers 7 to 10 and 12 adhere to the belt conveyor 22, they are disinfected by the disinfecting liquid 23 and discharged from the second space 4 to the first space 2 or the belt. There is an advantage that adhesion to other containers 7 to 10 and 12 via the conveyor 22 can be prevented. Further, since the sterilization lamp 17 is provided in the second space 4 and the ethanol shower 25 is provided in the pass boxes 14 and 15, the second space 4 to the first space 2 is provided. Release of dust and the like can be prevented more reliably.

In the present embodiment, the second space 4 is used as a criterion for opening / closing the pass boxes 14, 15 by the control unit 31 and a criterion for opening / closing the lids 7a-10a, 12a of the various containers 7-10, 12. Whether or not the cleanliness of the inside is class 100 or higher is adopted, but other cleanliness may be adopted as a criterion.
Further, in the present embodiment, the subculture operation has been described as an example, but it can be similarly applied to other processing such as a medium replacement operation instead.

  In addition, although the two pass boxes 14 and 15 are provided as the pass boxes 14 and 15 for introducing and taking out the various containers 7 to 10 and 12 and the chip box from the second space, the present invention is not limited to this. Instead, it may be one or three or more. It is preferable that a plurality of pass boxes 14 and 15 are provided in order to make the next container stand by and perform the culture process efficiently while introducing and removing one container.

It is a whole lineblock diagram showing the culture treatment device concerning one embodiment of this invention. It is a whole block diagram which shows the introduction process of the culture container and the chip box in the pass box by the culture processing apparatus of FIG. It is a whole block diagram explaining the chip | tip mounting process by the culture processing apparatus of FIG. It is a whole block diagram explaining the culture medium aspiration process by the culture processing apparatus of FIG. It is a whole block diagram explaining the culture medium discarding process by the culture processing apparatus of FIG. It is a whole block diagram explaining the chemical | medical solution suction process by the culture processing apparatus of FIG. It is a whole block diagram explaining the cell peeling process by the culture processing apparatus of FIG. It is a whole block diagram explaining the injection | pouring process of the cell suspension to the centrifuge container by the culture processing apparatus of FIG.

Explanation of symbols

1 Culture treatment device 4 Second space (small space)
7 Chemical solution containers 7a to 10a, 12a Lid 8 Serum container 9 Culture container 14, 15 Pass box 18 Particle counter (cleanness sensor)
DESCRIPTION OF SYMBOLS 19 Opening / closing mechanism 20 Dispensing apparatus 21 Conveying mechanism 22,22 'Belt conveyor 24 Disinfection liquid tank 31 Control part

Claims (4)

A culture treatment apparatus for performing a predetermined treatment on cells contained in a culture vessel,
A small space partitioned from the outside, an airlock-type pass box in which various containers such as a culture container, a drug solution container, and a serum container that can be opened and closed by a lid body can be taken in and out of the small space, and the pass box Equipped with a transport mechanism that can transport various containers into and out of a small space,
An opening / closing mechanism for opening and closing the lids of the various containers in the small space, a dispensing device capable of sucking and dispensing chemicals from the various containers introduced, and a cleanliness for measuring the cleanliness in the small space Sensor and
A culture treatment apparatus comprising a control unit that prohibits introduction of various containers into a small space when the cleanliness measured by the cleanliness sensor does not satisfy a predetermined cleanliness. The transport mechanism is provided in each of the pass box and the small space, and includes a plurality of belt conveyors that deliver various containers to be transported,
The culture treatment apparatus according to claim 1, wherein a disinfectant bath for immersing a part of the belt conveyor is provided at the bottom of the pass box and the small space. A culture treatment method for supplying liquid contained in various containers such as a drug solution container and a serum container to cells contained in a culture container,
Introducing various containers in a small space where the cleanliness is controlled,
Open the lids of various introduced containers,
Suction and hold the liquid inside various containers,
After closing the lids of various containers and taking them out of the small space,
Introduce a culture container in a small space,
Open the lid of the introduced culture vessel,
A culture treatment method for injecting liquids held in various containers into a culture container. After confirming that the cleanliness in the small space is equal to or greater than the specified cleanliness,
The culture treatment method according to claim 3, wherein a culture container and various containers are introduced into the small space.

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